Purity assignment for peptide certified reference materials by combining qNMR and LC-MS/MS amino acid analysis results: application to angiotensin II

Analytical and Bioanalytical Chemistry
Jeremy E MelansonJuris Meija

Abstract

The purity value assignment of metrologically traceable peptide reference standards requires specialized primary methods. Conventionally, amino acid analysis by isotope dilution tandem mass spectrometry (LC-MS/MS) following peptide hydrolysis is employed as a reference method. By contrast, quantitative nuclear magnetic resonance (qNMR) spectroscopy allows for quantitation of intact peptides, thus eliminating potential bias due to hydrolysis. Both methods are susceptible to interference from related peptide impurities, which need to be accurately measured and accounted for. The mass balance approach has also been employed for peptide purity measurements, whereby the purity is defined by the sum of the mass fraction of all impurities identified. Ideally, results from these three orthogonal methods can be combined for final purity assignment of peptide reference standards. Here we report a novel strategy for correcting both LC-MS/MS and 1H-qNMR results for related peptide impurities and combining results from both methods using a Bayesian statistical approach using mass balance results as prior knowledge. The mass balance method relied on a validated 19F-qNMR method to measure the trifluoroacetic acid (TFA) counter-ion, considered...Continue Reading

References

Jan 23, 1999·Journal of Chromatography. a·M Fountoulakis, H W Lahm
Nov 30, 2011·Methods in Molecular Biology·David M Bunk, Mark S Lowenthal
Feb 14, 2012·Analytical and Bioanalytical Chemistry·Torsten Schoenberger
Jul 20, 2013·Analytical Chemistry·Caroline PritchardAlison E Ashcroft
Oct 9, 2014·Journal of Medicinal Chemistry·Guido F PauliJosé G Napolitano
Sep 9, 2015·Analytical and Bioanalytical Chemistry·Michael A NelsonKatrice A Lippa
Sep 18, 2017·Analytical and Bioanalytical Chemistry·Juris Meija
Feb 17, 2018·Journal of the American Society for Mass Spectrometry·Bradley B Stocks, Jeremy E Melanson
Aug 22, 2018·Analytical and Bioanalytical Chemistry·Bradley B StocksJeremy E Melanson

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Citations

Apr 20, 2020·Natural Products and Bioprospecting·Dezhi YangGuanhua Du
Aug 22, 2018·Analytical and Bioanalytical Chemistry·Bradley B StocksJeremy E Melanson
Jan 15, 2019·Journal of Pharmaceutical and Biomedical Analysis·Chensheng LiFouad Atouf
Feb 23, 2019·The Journal of Organic Chemistry·Mary P ChoulesGuido F Pauli
Dec 11, 2020·Analytical Chemistry·Adilah BahadoorJeremy E Melanson

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